• Product name
    Anti-Amyloid Precursor Protein antibody [DE2B4]
    See all Amyloid Precursor Protein primary antibodies
  • Description
    Mouse monoclonal [DE2B4] to Amyloid Precursor Protein
  • Host species
  • Specificity
    Reacts with beta-amyloid peptide and secreted amyloid precursor protein (alpha-secretase cleaved). This antibody has very weak recognition of full length APP and we do not recommend it for this purpose. It does however strongly recognize alpha-secretase cleaved soluble APP as well as b-amyloid peptide (40 and 42), in both cases native an denatured forms.
  • Tested applications
    Suitable for: WB, ICC, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Cow, Human, Pig, Monkey
    Does not react with: Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human Amyloid Precursor Protein aa 672-689 conjugated to keyhole limpet haemocyanin. Corresponding to amino acids 1-16 of amyloid beta protein.

  • Epitope
    Ab12266 recognizes an epitope within amino acids 1-17 of the amyloid beta/A4 peptide region of APP. As such it recognises the C-terminus of alpha-secretase cleaved APP and amyloid beta/A4 peptide. It does not recognize the signal peptide of APP.
  • General notes
    Please note this antibody is very susceptible to freezing. Do NOT freeze.



Our Abpromise guarantee covers the use of ab12266 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/50 - 1/500.
ICC Use at an assay dependent concentration. Use ice-cold methanol fixation for 10 minutes, followed by a brief wash with TBS. We have also used 4% paraformaldehyde for 20 minutes followed by permeabilization with either methanol of 0.5% Triton X-100 in TBS and this also seems to work well.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Retrieve antigens with 70% Formic acid for 10-30 minutes at room temperature before commencing with IHC staining protocol. Please note: it is important if ab12266 is being used to stain amyloid plaques in Alzheimer's disease brains that the sections are pre-treated with formic acid. This requirement applies to most, if not all amyloid antibodies and is not unique to this product.


  • Function
    Functions as a cell surface receptor and performs physiological functions on the surface of neurons relevant to neurite growth, neuronal adhesion and axonogenesis. Involved in cell mobility and transcription regulation through protein-protein interactions. Can promote transcription activation through binding to APBB1-KAT5 and inhibits Notch signaling through interaction with Numb. Couples to apoptosis-inducing pathways such as those mediated by G(O) and JIP. Inhibits G(o) alpha ATPase activity (By similarity). Acts as a kinesin I membrane receptor, mediating the axonal transport of beta-secretase and presenilin 1. Involved in copper homeostasis/oxidative stress through copper ion reduction. In vitro, copper-metallated APP induces neuronal death directly or is potentiated through Cu(2+)-mediated low-density lipoprotein oxidation. Can regulate neurite outgrowth through binding to components of the extracellular matrix such as heparin and collagen I and IV. The splice isoforms that contain the BPTI domain possess protease inhibitor activity. Induces a AGER-dependent pathway that involves activation of p38 MAPK, resulting in internalization of amyloid-beta peptide and leading to mitochondrial dysfunction in cultured cortical neurons. Provides Cu(2+) ions for GPC1 which are required for release of nitric oxide (NO) and subsequent degradation of the heparan sulfate chains on GPC1.
    Beta-amyloid peptides are lipophilic metal chelators with metal-reducing activity. Bind transient metals such as copper, zinc and iron. In vitro, can reduce Cu(2+) and Fe(3+) to Cu(+) and Fe(2+), respectively. Beta-amyloid 42 is a more effective reductant than beta-amyloid 40. Beta-amyloid peptides bind to lipoproteins and apolipoproteins E and J in the CSF and to HDL particles in plasma, inhibiting metal-catalyzed oxidation of lipoproteins. Beta-APP42 may activate mononuclear phagocytes in the brain and elicit inflammatory responses. Promotes both tau aggregation and TPK II-mediated phosphorylation. Interaction with overexpressed HADH2 leads to oxidative stress and neurotoxicity. Also binds GPC1 in lipid rafts.
    Appicans elicit adhesion of neural cells to the extracellular matrix and may regulate neurite outgrowth in the brain.
    The gamma-CTF peptides as well as the caspase-cleaved peptides, including C31, are potent enhancers of neuronal apoptosis.
    N-APP binds TNFRSF21 triggering caspase activation and degeneration of both neuronal cell bodies (via caspase-3) and axons (via caspase-6).
  • Tissue specificity
    Expressed in all fetal tissues examined with highest levels in brain, kidney, heart and spleen. Weak expression in liver. In adult brain, highest expression found in the frontal lobe of the cortex and in the anterior perisylvian cortex-opercular gyri. Moderate expression in the cerebellar cortex, the posterior perisylvian cortex-opercular gyri and the temporal associated cortex. Weak expression found in the striate, extra-striate and motor cortices. Expressed in cerebrospinal fluid, and plasma. Isoform APP695 is the predominant form in neuronal tissue, isoform APP751 and isoform APP770 are widely expressed in non-neuronal cells. Isoform APP751 is the most abundant form in T-lymphocytes. Appican is expressed in astrocytes.
  • Involvement in disease
    Alzheimer disease 1
    Cerebral amyloid angiopathy, APP-related
  • Sequence similarities
    Belongs to the APP family.
    Contains 1 BPTI/Kunitz inhibitor domain.
  • Domain
    The basolateral sorting signal (BaSS) is required for sorting of membrane proteins to the basolateral surface of epithelial cells.
    The NPXY sequence motif found in many tyrosine-phosphorylated proteins is required for the specific binding of the PID domain. However, additional amino acids either N- or C-terminal to the NPXY motif are often required for complete interaction. The PID domain-containing proteins which bind APP require the YENPTY motif for full interaction. These interactions are independent of phosphorylation on the terminal tyrosine residue. The NPXY site is also involved in clathrin-mediated endocytosis.
  • Post-translational
    Proteolytically processed under normal cellular conditions. Cleavage either by alpha-secretase, beta-secretase or theta-secretase leads to generation and extracellular release of soluble APP peptides, S-APP-alpha and S-APP-beta, and the retention of corresponding membrane-anchored C-terminal fragments, C80, C83 and C99. Subsequent processing of C80 and C83 by gamma-secretase yields P3 peptides. This is the major secretory pathway and is non-amyloidogenic. Alternatively, presenilin/nicastrin-mediated gamma-secretase processing of C99 releases the amyloid beta proteins, amyloid-beta 40 (Abeta40) and amyloid-beta 42 (Abeta42), major components of amyloid plaques, and the cytotoxic C-terminal fragments, gamma-CTF(50), gamma-CTF(57) and gamma-CTF(59). Many other minor beta-amyloid peptides, beta-amyloid 1-X peptides, are found in cerebral spinal fluid (CSF) including the beta-amyloid X-15 peptides, produced from the cleavage by alpha-secretase and all terminating at Gln-686.
    Proteolytically cleaved by caspases during neuronal apoptosis. Cleavage at Asp-739 by either caspase-6, -8 or -9 results in the production of the neurotoxic C31 peptide and the increased production of beta-amyloid peptides.
    N- and O-glycosylated. O-glycosylation on Ser and Thr residues with core 1 or possibly core 8 glycans. Partial tyrosine glycosylation (Tyr-681) is found on some minor, short beta-amyloid peptides (beta-amyloid 1-15, 1-16, 1-17, 1-18, 1-19 and 1-20) but not found on beta-amyloid 38, beta-amyloid 40 nor on beta-amyloid 42. Modification on a tyrosine is unusual and is more prevelant in AD patients. Glycans had Neu5AcHex(Neu5Ac)HexNAc-O-Tyr, Neu5AcNeu5AcHex(Neu5Ac)HexNAc-O-Tyr and O-AcNeu5AcNeu5AcHex(Neu5Ac)HexNAc-O-Tyr structures, where O-Ac is O-acetylation of Neu5Ac. Neu5AcNeu5Ac is most likely Neu5Ac 2,8Neu5Ac linked. O-glycosylations in the vicinity of the cleavage sites may influence the proteolytic processing. Appicans are L-APP isoforms with O-linked chondroitin sulfate.
    Phosphorylation in the C-terminal on tyrosine, threonine and serine residues is neuron-specific. Phosphorylation can affect APP processing, neuronal differentiation and interaction with other proteins. Phosphorylated on Thr-743 in neuronal cells by Cdc5 kinase and Mapk10, in dividing cells by Cdc2 kinase in a cell-cycle dependent manner with maximal levels at the G2/M phase and, in vitro, by GSK-3-beta. The Thr-743 phosphorylated form causes a conformational change which reduces binding of Fe65 family members. Phosphorylation on Tyr-757 is required for SHC binding. Phosphorylated in the extracellular domain by casein kinases on both soluble and membrane-bound APP. This phosphorylation is inhibited by heparin.
    Extracellular binding and reduction of copper, results in a corresponding oxidation of Cys-144 and Cys-158, and the formation of a disulfide bond. In vitro, the APP-Cu(+) complex in the presence of hydrogen peroxide results in an increased production of beta-amyloid-containing peptides.
    Trophic-factor deprivation triggers the cleavage of surface APP by beta-secretase to release sAPP-beta which is further cleaved to release an N-terminal fragment of APP (N-APP).
    Beta-amyloid peptides are degraded by IDE.
  • Cellular localization
    Membrane. Membrane, clathrin-coated pit. Cell surface protein that rapidly becomes internalized via clathrin-coated pits. During maturation, the immature APP (N-glycosylated in the endoplasmic reticulum) moves to the Golgi complex where complete maturation occurs (O-glycosylated and sulfated). After alpha-secretase cleavage, soluble APP is released into the extracellular space and the C-terminal is internalized to endosomes and lysosomes. Some APP accumulates in secretory transport vesicles leaving the late Golgi compartment and returns to the cell surface. Gamma-CTF(59) peptide is located to both the cytoplasm and nuclei of neurons. It can be translocated to the nucleus through association with APBB1 (Fe65). Beta-APP42 associates with FRPL1 at the cell surface and the complex is then rapidly internalized. APP sorts to the basolateral surface in epithelial cells. During neuronal differentiation, the Thr-743 phosphorylated form is located mainly in growth cones, moderately in neurites and sparingly in the cell body. Casein kinase phosphorylation can occur either at the cell surface or within a post-Golgi compartment. Associates with GPC1 in perinuclear compartments. Colocalizes with SORL1 in a vesicular pattern in cytoplasm and perinuclear regions.
  • Information by UniProt
  • Database links
  • Alternative names
    • A4 amyloid protein antibody
    • A4_HUMAN antibody
    • AAA antibody
    • ABETA antibody
    • ABPP antibody
    • AD1 antibody
    • AICD-50 antibody
    • AICD-57 antibody
    • AICD-59 antibody
    • AID(50) antibody
    • AID(57) antibody
    • AID(59) antibody
    • Alzheimer disease amyloid protein antibody
    • Amyloid beta (A4) precursor protein antibody
    • Amyloid beta A4 protein antibody
    • Amyloid beta protein antibody
    • Amyloid intracellular domain 50 antibody
    • Amyloid intracellular domain 57 antibody
    • Amyloid intracellular domain 59 antibody
    • Amyloid precursor protein antibody
    • APP antibody
    • APPI antibody
    • Beta amyloid peptide antibody
    • Beta-amyloid precursor protein antibody
    • Beta-APP40 antibody
    • Beta-APP42 antibody
    • C31 antibody
    • Cerebral vascular amyloid peptide antibody
    • CTFgamma antibody
    • CVAP antibody
    • Gamma-CTF(50) antibody
    • Gamma-CTF(57) antibody
    • Gamma-CTF(59) antibody
    • peptidase nexin II antibody
    • PN 2 antibody
    • PN II antibody
    • PN-II antibody
    • PN2 antibody
    • PreA4 antibody
    • Protease nexin II antibody
    • Protease nexin-II antibody
    • S-APP-alpha antibody
    • S-APP-beta antibody
    see all


  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Amyloid Precursor Protein knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HepG2 whole cell lysate (20 µg)
    Lane 4: HeLa whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab12266 observed at 110 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab12266 was shown to specifically react with Amyloid Precursor Protein when Amyloid Precursor Protein knockout samples were used. Wild-type and Amyloid Precursor Protein knockout samples were subjected to SDS-PAGE. ab12266 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/50 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry of formaldehyde fixed brain tissue, after antigen-retrieval by formic acid treatment, for post-mortem diagnosis of Alzheimer s disease.
  • A section from the temporal lobe of a patient who died with Alzheimer’s disease was pre-treated with formic acid for 10 minutes prior to reaction with mouse monoclonal antibody DE4B2 (ab12266) that recognises an epitope near the N-terminus of β-amyloid peptides.  Bound antibody was detected by reaction with alkaline phosphatase-conjugated goat ant mouse IgG followed by BCIP/NBT substrate.  Plaques in the neuropil have been circled in black. Amyloid deposits stained by DE2B4 (ab12266) can be seen around some blood vessels (*) but some are not stained (#)
  • IHC image of ab12266 staining in human Alzheimer brain (cerebral cortex) formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab12266, at a 1 in 40 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.


This product has been referenced in:
  • Kim J  et al. HIV-1 Tat Interacts with and Regulates the Localization and Processing of Amyloid Precursor Protein. PLoS One 8:e77972 (2013). IP ; Human . Read more (PubMed: 24312169) »
  • Fatemi SH  et al. Impairment of fragile X mental retardation protein-metabotropic glutamate receptor 5 signaling and its downstream cognates ras-related C3 botulinum toxin substrate 1, amyloid beta A4 precursor protein, striatal-enriched protein tyrosine phosphatase, and homer 1, in autism: a postmortem study in cerebellar vermis and superior frontal cortex. Mol Autism 4:21 (2013). WB ; Human . Read more (PubMed: 23803181) »
See all 5 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A


Thank you for contacting us. A collaborator has used ellipsometry to measure the affinity of this antibody for a synthetic peptide corresponding to amino acids 1-16 of beta peptide and gets a value of around 10-8 M. This is a very preliminary measurement, but it may provide a starting point for your technique. I hope this helps, please let me know if you need any additional information.

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I received the following information from the researcher who tested the antibody ab12266 in ICC: "We routinely use ice-cold methanol fixation for 10 minutes, followed by a brief wash with TBS. We have also used 4% paraformaldehyde for 20 minutes followed by permeabilization with either methanol of 0.5% Triton X-100 in TBS and this also seems to work fine." Please let me know if you need further assistance,

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BATCH NUMBER 176737 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE I have transfected the DNAs encoding either APP or factor A into human cancer cells. The number of cell was counted and resuspended in appropriate amount of 2X SDS-PAGE sample buffer and boiled for 3min. PRIMARY ANTIBODY The membrane was blocked with 5% skim milk in TBS-T buffer. Anti-APP antibody was diluted at first 1:500 and later 1:100 and added to the membrane and incubated overnight with gentle rocking. The membrane was incubated further 30min at RT, washed three times with TBS-T DETECTION METHOD After 3 times washing (10min for each washing), ECL solution (Amersham-Pharmacia) was added, incubated for 1 min and the membrane was developed with Hyperfilm (Amersham-Pharmacia) by exposing the film for 5 min, 20 min, 2 hours, or overnight POSITIVE AND NEGATIVE CONTROLS USED I also tried to detect endogenous APP in the same cell line by using another company's antibody and there was no problem for detection. ANTIBODY STORAGE CONDITIONS 4C ELECTROPHORESIS/GEL CONDITIONS 8% SDS-PAGE TRANSFER AND BLOCKING CONDITIONS transferred activated PVDF membrane for 90 min at 60V SECONDARY ANTIBODY secondary antibody (anti-mouse) was added and incubated for 2 hours. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? See above ADDITIONAL NOTES While i was filling the complaint report, i repeated the experiment with fresh primary and secondary antibody and i still could not get the target band. There were several faint background bands but no APP specific band.

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Thank you for your enquiry. I am sorry to hear that your customer has been having difficulties using this antibody by western blotting. Their approach is largely on that I would recommend. They have tried various dilutions using an overnight incubation. I am certainly prepared to offer your customer credit if the antibody was purchased within the past 90 days. If this is the case please e-mail me details of the order including the date of purchase and order number.

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Thank you for your enquiry and your interest in our products. Ab12266 and ab10146 are from two different clones and sources. They have been tested for different applications. For further information, please take a look at the on-line datasheets and decide which one would suit your need best.

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